Detection, Quantification, and Isomer Differentiation of Per- and Polyfluoroalkyl Substances (PFAS) Using MALDI-TOF with Trapped Ion Mobility.

Per- and polyfluoroalkyl substances (PFAS) are a class of organic compounds that have attracted global attention for their persistence in the environment, exposure to biological organisms, and their adverse health effects. There is an urgent need to develop analytical methodologies for the characterization of PFAS in various sample matrices. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) represents a chromatography-free MS method that performs laser-based ionization and in situ analysis on samples. In this study, we present PFAS analysis by MALDI-time-of-flight (TOF) MS with trapped ion mobility spectrometry (TIMS), which provides an additional dimension of gas phase separation based on the size-to-charge ratios. MALDI matrix composition and key instrument parameters were optimized to produce different ranges of calibration curves. Parts per billion (ppb) range of calibration curves were achieved for a list of legacy and alternative perfluorosulfonic acids (PFSAs) and perfluorocarboxylic acids (PFCAs), while ion mobility spectrum filtering enabled parts per trillion (ppt) range of calibration curves for PFSAs. We also successfully demonstrated the separation of three perfluorooctanesulfonic acid (PFOS) structural isomers in the gas phase using TIMS. Our results demonstrated the new development of utilizing MALDI-TOF-MS coupled with TIMS for fast, quantitative, and sensitive analysis of PFAS, paving ways to future high-throughput and in situ analysis of PFAS such as MS imaging applications.

Biodistribution and racemization of gut-absorbed L/D-alanine in germ-free mice.

Microbiome-derived metabolites are important for the microbiome-gut-brain axis and the discovery of new disease treatments. D-Alanine (D-Ala) is found in many animals as a potential co-agonist of the N-methyl-D-aspartate receptors (NMDAR), receptors widely used in the nervous and endocrine systems. The gut microbiome, diet and putative endogenous synthesis are the potential sources of D-Ala in animals, although there is no direct evidence to show the distribution and racemization of gut-absorbed L-/D-Ala with regards to host-microbe interactions in mammals. In this work, we utilized germ-free mice to control the interference from microbiota and isotopically labeled L-/D-Ala to track their biodistribution and racemization in vivo. Results showed time-dependent biodistribution of gut-absorbed D-Ala, particularly accumulation of gut-absorbed D-Ala in pancreatic tissues, brain, and pituitary. No endogenous synthesis of D-Ala via racemization was observed in germ-free mice. The sources of D-Ala in mice were revealed as microbiota and diet, but not endogenous racemization. This work indicates the importance of further investigating the in vivo biological functions of gut-microbiome derived D-Ala, particularly on NMDAR-related activities, for D-Ala as a potential signaling molecules in the microbiome-gut-brain axis.